Title:
ENGINEERED GUIDE RNA COMPRISING U-RICH TAIL FOR INCREASING EFFICIENCY OF CRISPR/CAS12F1 SYSTEM, AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2022/075808
Kind Code:
A1
Abstract:
The present specification provides an engineered Cas12f1 guide RNA that overcomes limitations of a conventional technology in order to enhance the intracellular gene editing efficiency of a CRISPR/Cas12f1 system. The engineered Cas12f1 guide RNA is obtained by adding a new element to a guide RNA found in nature, and a part of the structure thereof is modified. The engineered Cas12f1 guide RNA comprises, at the 3' terminus of the construct, a U-rich tail, which is a new element. In addition, at least a part of a scaffold region capable of interacting with the Cas12f1 protein is modified in the formation of the engineered Cas12f1 guide RNA.
Inventors:
KIM YONG-SAM (KR)
KIM DO YON (KR)
KIM DO YON (KR)
Application Number:
PCT/KR2021/013898
Publication Date:
April 14, 2022
Filing Date:
October 08, 2021
Export Citation:
Assignee:
GENKORE INC (KR)
International Classes:
C12N9/22; C12N15/10; C12N15/113; C12N15/63; C12N15/90
Domestic Patent References:
| WO2019089820A1 | 2019-05-09 |
Foreign References:
| US20200172886A1 | 2020-06-04 | |||
| CN111363763A | 2020-07-03 | |||
| US20200190494A1 | 2020-06-18 | |||
| US20200190494A1 | 2020-06-18 | |||
| KR2020014961W | 2020-10-29 |
Other References:
MOON, S. B.: "Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang", NATURE COMMUNICATIONS, vol. 9, 2018, pages 1 - 11, XP055785455, DOI: 10.1038/s41467-018-06129-w
KARVELIS TAUTVYDAS, BIGELYTE GRETA, YOUNG JOSHUA K, HOU ZHENGLIN, ZEDAVEINYTE RIMANTE, BUDRE KAROLINA, PAULRAJ SUSHMITHA, DJUKANOV: "PAM recognition by miniature CRISPR–Cas12f nucleases triggers programmable double-stranded DNA target cleavage", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, GB, vol. 48, no. 9, 21 May 2020 (2020-05-21), GB , pages 5016 - 5023, XP055920188, ISSN: 0305-1048, DOI: 10.1093/nar/gkaa208
HARRINGTON, L. B.: "Programmed DNA destruction by miniature CRISPR-Cas14 enzymes", SCIENCE, vol. 362, 18 August 1612 (1612-08-18), pages 839 - 842, XP055614750, DOI: 10.1126/science.aav4294
KIM DO YON; LEE JEONG MI; MOON SU BIN; CHIN HYUN JUNG; PARK SEYEON; LIM YOUJUNG; KIM DAESIK; KOO TAEYOUNG; KO JEONG-HEON; KIM YONG: "Efficient CRISPR editing with a hypercompact Cas12f1 and engineered guide RNAs delivered by adeno-associated virus", NATURE BIOTECHNOLOGY, NATURE PUBLISHING GROUP US, NEW YORK, vol. 40, no. 1, 2 September 2021 (2021-09-02), New York, pages 94 - 102, XP037667066, ISSN: 1087-0156, DOI: 10.1038/s41587-021-01009-z
HARRINGTON ET AL.: "Programmed DNA destruction by miniature CRISPR-Cas14 enzymes", SCIENCE, vol. 362, 2018, pages 839 - 842, XP055614750, DOI: 10.1126/science.aav4294
KARVELIS ET AL., NUCLEIC ACIDS RESEARCH, vol. 48, no. 9, 2020, pages 5017
TAKEDA ET AL.: "Structure of the miniature type V-F CRISPR-Cas effector enzyme", MOLECULAR CELL, vol. 81, 2021, pages 1 - 13
XIAO ET AL.: "Structural basis for the dimerization-dependent CRISPR-Cas12f nuclease", BIORXIV, 2020
MAKAROVA ET AL.: "Nature Reviews", MICROBIOLOGY, vol. 18, 2020, pages 67
MIYAGISHI ET AL., NATURE BIOTECHNOLOGY, vol. 20, 2002, pages 497 - 500
XIA ET AL., NUCLEIC ACIDS RES., vol. 1, no. 17, September 2003 (2003-09-01), pages 31
PANYAM, ADV DRUG DELIV REV, 13 September 2012 (2012-09-13)
KARVELIS TAUTVYDAS, BIGELYTE GRETA, YOUNG JOSHUA K, HOU ZHENGLIN, ZEDAVEINYTE RIMANTE, BUDRE KAROLINA, PAULRAJ SUSHMITHA, DJUKANOV: "PAM recognition by miniature CRISPR–Cas12f nucleases triggers programmable double-stranded DNA target cleavage", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, GB, vol. 48, no. 9, 21 May 2020 (2020-05-21), GB , pages 5016 - 5023, XP055920188, ISSN: 0305-1048, DOI: 10.1093/nar/gkaa208
HARRINGTON, L. B.: "Programmed DNA destruction by miniature CRISPR-Cas14 enzymes", SCIENCE, vol. 362, 18 August 1612 (1612-08-18), pages 839 - 842, XP055614750, DOI: 10.1126/science.aav4294
KIM DO YON; LEE JEONG MI; MOON SU BIN; CHIN HYUN JUNG; PARK SEYEON; LIM YOUJUNG; KIM DAESIK; KOO TAEYOUNG; KO JEONG-HEON; KIM YONG: "Efficient CRISPR editing with a hypercompact Cas12f1 and engineered guide RNAs delivered by adeno-associated virus", NATURE BIOTECHNOLOGY, NATURE PUBLISHING GROUP US, NEW YORK, vol. 40, no. 1, 2 September 2021 (2021-09-02), New York, pages 94 - 102, XP037667066, ISSN: 1087-0156, DOI: 10.1038/s41587-021-01009-z
HARRINGTON ET AL.: "Programmed DNA destruction by miniature CRISPR-Cas14 enzymes", SCIENCE, vol. 362, 2018, pages 839 - 842, XP055614750, DOI: 10.1126/science.aav4294
KARVELIS ET AL., NUCLEIC ACIDS RESEARCH, vol. 48, no. 9, 2020, pages 5017
TAKEDA ET AL.: "Structure of the miniature type V-F CRISPR-Cas effector enzyme", MOLECULAR CELL, vol. 81, 2021, pages 1 - 13
XIAO ET AL.: "Structural basis for the dimerization-dependent CRISPR-Cas12f nuclease", BIORXIV, 2020
MAKAROVA ET AL.: "Nature Reviews", MICROBIOLOGY, vol. 18, 2020, pages 67
MIYAGISHI ET AL., NATURE BIOTECHNOLOGY, vol. 20, 2002, pages 497 - 500
XIA ET AL., NUCLEIC ACIDS RES., vol. 1, no. 17, September 2003 (2003-09-01), pages 31
PANYAM, ADV DRUG DELIV REV, 13 September 2012 (2012-09-13)
Attorney, Agent or Firm:
IPLAY PATENT LAW FIRM (KR)
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